title: A comparative study of matrix metalloproteinase and aggrecanase mediated release of latent cytokines at arthritic joints.
abstract: BACKGROUND
  Latent cytokines are engineered by fusing the latency associated peptide (LAP) derived from transforming growth factor-β (TGF-β) with the therapeutic cytokine, in this case interferon-β (IFN-β), via an inflammation-specific matrix metalloproteinase (MMP) cleavage site.
  
  
  OBJECTIVE
  To demonstrate latency and specific delivery in vivo and to compare therapeutic efficacy of aggrecanase-mediated release of latent IFN-β in arthritic joints to the original MMP-specific release.
  
  
  METHODS
  Recombinant fusion proteins with MMP, aggrecanase or devoid of cleavage site were expressed in CHO cells, purified and characterised in vitro by Western blotting and anti-viral protection assays. Therapeutic efficacy and half-life were assessed in vivo using the mouse collagen-induced arthritis model (CIA) of rheumatoid arthritis and a model of acute paw inflammation, respectively. Transgenic mice with an IFN-regulated luciferase gene were used to assess latency in vivo and targeted delivery to sites of disease.
  
  
  RESULTS
  Efficient localised delivery of IFN-β to inflamed paws, with low levels of systemic delivery, was demonstrated in transgenic mice using latent IFN-β. Engineering of latent IFN-β with an aggrecanase-sensitive cleavage site resulted in efficient cleavage by ADAMTS-4, ADAMTS-5 and synovial fluid from arthritic patients, with an extended half-life similar to the MMP-specific molecule and greater therapeutic efficacy in the CIA model.
  
  
  CONCLUSIONS
  Latent cytokines require cleavage in vivo for therapeutic efficacy, and they are delivered in a dose dependent fashion only to arthritic joints. The aggrecanase-specific cleavage site is a viable alternative to the MMP cleavage site for the targeting of latent cytokines to arthritic joints.
authors.affiliation: Bone and Joint Research Unit, William Harvey Research Institute, Queen Mary University of London, London, UK.
authors.forename: Lisa
authors.initials: L
authors.lastname: Mullen
authors.affiliation: Bone and Joint Research Unit, William Harvey Research Institute, Queen Mary University of London, London, UK.
authors.forename: Gill
authors.initials: G
authors.lastname: Adams
authors.affiliation: Department of Molecular Oncology, Barts Cancer Institute, Queen Mary University of London, London, UK.
authors.forename: Julie
authors.initials: J
authors.lastname: Foster
authors.affiliation: Bone and Joint Research Unit, William Harvey Research Institute, Queen Mary University of London, London, UK.
authors.forename: Sandrine
authors.initials: S
authors.lastname: Vessillier
authors.affiliation: Department of Gene Regulation and Differentiation, Helmholtz Centre for Infection Research, Braunschweig, Germany.
authors.forename: Mario
authors.initials: M
authors.lastname: Köster
authors.affiliation: Department of Gene Regulation and Differentiation, Helmholtz Centre for Infection Research, Braunschweig, Germany.
authors.forename: Hansjörg
authors.initials: H
authors.lastname: Hauser
authors.affiliation: Bone and Joint Research Unit, William Harvey Research Institute, Queen Mary University of London, London, UK.
authors.forename: Lorna
authors.initials: L
authors.lastname: Layward
authors.affiliation: Bone and Joint Research Unit, William Harvey Research Institute, Queen Mary University of London, London, UK.
authors.forename: David
authors.initials: D
authors.lastname: Gould
authors.affiliation: Bone and Joint Research Unit, William Harvey Research Institute, Queen Mary University of London, London, UK.
authors.forename: Yuti
authors.initials: Y
authors.lastname: Chernajovsky
mesh_terms: D000818:Animals; D018501:Antirheumatic Agents / Q000008:administration & dosage* / Q000493:pharmacokinetics / Q000627:therapeutic use; D001169:Arthritis, Experimental / Q000188:drug therapy*; D001172:Arthritis, Rheumatoid / Q000188:drug therapy*; D016466:CHO Cells; D003412:Cricetulus; D016207:Cytokines; D004305:Dose-Response Relationship, Drug; D016503:Drug Delivery Systems / Q000379:methods; D004353:Drug Evaluation, Preclinical / Q000379:methods; D010450:Endopeptidases; D006207:Half-Life; D016899:Interferon-beta / Q000008:administration & dosage* / Q000493:pharmacokinetics / Q000627:therapeutic use; D020782:Matrix Metalloproteinases; D051379:Mice; D008822:Mice, Transgenic; D011993:Recombinant Fusion Proteins / Q000008:administration & dosage / Q000493:pharmacokinetics / Q000627:therapeutic use
chemical_list: D018501:Antirheumatic Agents; D016207:Cytokines; D011993:Recombinant Fusion Proteins; D016899:Interferon-beta; D010450:Endopeptidases; D020782:Matrix Metalloproteinases; C083641:aggrecanase
keywords: Cytokines; Pharmacokinetics; Rheumatoid Arthritis; Treatment